                         SEQUENCE LISTING

<110>  Duke University
       Heaton, Nicholas
       Harding, Alfred
       Heaton, Brook
       Chaparian, Ryan
 
<120>  ENGINEERED INFLUENZA VIRUSES EXPRESSING SARS-COV-2 ANTIGENS, 
       VACCINES AND METHODS OF MAKING AND USING THE SAME

<130>  155554.00645

<150>  63/180,472
<151>  2021-04-27

<160>  23    

<170>  PatentIn version 3.5

<210>  1
<211>  2736
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - engineered polypeptide derived from influenza A virus
       segment 4

<400>  1
agcaaaagca ggggaaaata aaaacaacca aattgaaggc aaacctactg gtcctgttaa       60

gtgcacttgc agctgcagtt gcagacacaa tttgtatagg ccaccatgaa ccccaatcaa      120

aaaattacaa ccatcgggtc tatctgcctc gtagtcggac tgatatccct aattcttcag      180

ataggtaata tcatatctat ctggatcagt catagtatac agactagagt gcagccaaca      240

gagtcaatcg tgagattccc aaatatcaca aatctgtgcc cattcggaga agtgttcaat      300

gcaacaagat tcgcatcagt gtacgcatgg aacagaaaga gaatctccaa ttgcgtggca      360

gactactcag tgctgtacaa ttcagcatca ttctcaacat tcaaatgcta cggagtgtca      420

ccaacaaaac ttaatgatct ttgcttcaca aatgtgtatg cagattcatt cgtgataaga      480

ggagatgaag tgagacaaat agcaccagga caaacaggaa aaatagcaga ttataattat      540

aaacttccag atgatttcac agggtgcgtg atcgcatgga attcaaacaa tctggactca      600

aaagtgggag ggaattacaa ctacctgtat agactgttca gaaaatcaaa tctcaaacca      660

ttcgaaagag acatctcaac agaaatctac caagcaggat caacaccatg caatggagtg      720

gaaggattca attgctattt cccactgcag tcatatggat tccagccaac aaatggagtg      780

ggatatcaac catatagagt ggtggtgctt tcattcgaac ttcttcatgc accagcaaca      840

gtgtgcggac caaaaaaatc aacaaatctt gtgaaaaata aatgcgtgaa tttcggcagt      900

ggggactata aggacgatga tgacaaagga tctggggcta ccaacttcag tctcctcaaa      960

caggccggag acgtggaaga aaatcctggg cctatgaaag cgaatttgtt agttttactg     1020

tccgcgttgg cggccgcgga cgcagacaca atatgtatag gctaccatgc gaacaattca     1080

accgacactg ttgacacagt actcgagaag aatgtgacag tgacacactc tgttaacctg     1140

ctcgaagaca gccacaacgg aaaactatgt agattaaaag gaatagcccc actacaattg     1200

gggaaatgta acatcgccgg atggctcttg ggaaacccag aatgcgaccc actgcttcca     1260

gtgagatcat ggtcctacat tgtagaaaca ccaaactctg agaatggaat atgttatcca     1320

ggagatttca tcgactatga ggagctgagg gagcaattga gctcagtgtc atcattcgaa     1380

agattcgaaa tatttcccaa agaaagctca tggcccaacc acaacacaaa cggagtaacg     1440

gcagcatgct cccatgaggg gaaaagcagt ttttacagaa atttgctatg gctgacggag     1500

aaggagggct catacccaaa gctgaaaaat tcttatgtga acaaaaaagg gaaagaagtc     1560

cttgtactgt ggggtattca tcacccgcct aacagtaagg aacaacagaa tctctatcag     1620

aatgaaaatg cttatgtctc tgtagtgact tcaaattata acaggagatt taccccggaa     1680

atagcagaaa gacccaaagt aagagatcaa gctgggagga tgaactatta ctggaccttg     1740

ctaaaacccg gagacacaat aatatttgag gcaaatggaa atctaatagc accaatgtat     1800

gctttcgcac tgagtagagg ctttgggtcc ggcatcatca cctcaaacgc atcaatgcat     1860

gagtgtaaca cgaagtgtca aacacccctg ggagctataa acagcagtct cccttaccag     1920

aatatacacc cagtcacaat aggagagtgc ccaaaatacg tcaggagtgc caaattgagg     1980

atggttacag gactaaggaa cactccgtcc attcaatcca gaggtctatt tggagccatt     2040

gccggtttta ttgaaggggg atggactgga atgatagatg gatggtatgg ttatcatcat     2100

cagaatgaac agggatcagg ctatgcagcg gatcaaaaaa gcacacaaaa tgccattaac     2160

gggattacaa acaaggtgaa cactgttatc gagaaaatga acattcaatt cacagctgtg     2220

ggtaaagaat tcaacaaatt agaaaaaagg atggaaaatt taaataaaaa agttgatgat     2280

ggatttctgg acatttggac atataatgca gaattgttag ttctactgga aaatgaaagg     2340

actctggatt tccatgactc aaatgtgaag aatctgtatg agaaagtaaa aagccaatta     2400

aagaataatg ccaaagaaat cggaaatgga tgttttgagt tctaccacaa gtgtgacaat     2460

gaatgcatgg aaagtgtaag aaatgggact tatgattatc ccaaatattc agaagagtca     2520

aagttgaaca gggaaaaggt agatggagtg aaattggaat caatggggat ctatcagatt     2580

ctggcgatct actcaactgt cgccagttca ctggtgcttt tggtctccct gggggcaatc     2640

agtttctgga tgtgttctaa tggatctttg cagtgcagaa tatgcatctg agattagaat     2700

ttcagaaata tgaggaaaaa cacccttgtt tctact                               2736


<210>  2
<211>  32
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - viral 5' UTR

<400>  2
agcaaaagca ggggaaaata aaaacaacca aa                                     32


<210>  3
<211>  67
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - viral packaging signal

<400>  3
ttgaaggcaa acctactggt cctgttaagt gcacttgcag ctgcagttgc agacacaatt       60

tgtatag                                                                 67


<210>  4
<211>  120
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - portion of a neuraminidase (NA) protein comprising 
       the transmembrane domain and cytoplasmic tail

<400>  4
atgaacccca atcaaaaaat tacaaccatc gggtctatct gcctcgtagt cggactgata       60

tccctaattc ttcagatagg taatatcata tctatctgga tcagtcatag tatacagact      120


<210>  5
<211>  669
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - receptor binding domain (RBD) of the spike protein 
       from the SARS-CoV-2 Wuhan variant

<400>  5
agagtgcagc caacagagtc aatcgtgaga ttcccaaata tcacaaatct gtgcccattc       60

ggagaagtgt tcaatgcaac aagattcgca tcagtgtacg catggaacag aaagagaatc      120

tccaattgcg tggcagacta ctcagtgctg tacaattcag catcattctc aacattcaaa      180

tgctacggag tgtcaccaac aaaacttaat gatctttgct tcacaaatgt gtatgcagat      240

tcattcgtga taagaggaga tgaagtgaga caaatagcac caggacaaac aggaaaaata      300

gcagattata attataaact tccagatgat ttcacagggt gcgtgatcgc atggaattca      360

aacaatctgg actcaaaagt gggagggaat tacaactacc tgtatagact gttcagaaaa      420

tcaaatctca aaccattcga aagagacatc tcaacagaaa tctaccaagc aggatcaaca      480

ccatgcaatg gagtggaagg attcaattgc tatttcccac tgcagtcata tggattccag      540

ccaacaaatg gagtgggata tcaaccatat agagtggtgg tgctttcatt cgaacttctt      600

catgcaccag caacagtgtg cggaccaaaa aaatcaacaa atcttgtgaa aaataaatgc      660

gtgaatttc                                                              669


<210>  6
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - flag tag

<400>  6
ggcagtgggg actataagga cgatgatgac aaa                                    33


<210>  7
<211>  66
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - 2A self-cleaving peptide

<400>  7
ggatctgggg ctaccaactt cagtctcctc aaacaggccg gagacgtgga agaaaatcct       60

gggcct                                                                  66


<210>  8
<211>  1698
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - influenza hemagglutinin (HA) protein

<400>  8
atgaaagcga atttgttagt tttactgtcc gcgttggcgg ccgcggacgc agacacaata       60

tgtataggct accatgcgaa caattcaacc gacactgttg acacagtact cgagaagaat      120

gtgacagtga cacactctgt taacctgctc gaagacagcc acaacggaaa actatgtaga      180

ttaaaaggaa tagccccact acaattgggg aaatgtaaca tcgccggatg gctcttggga      240

aacccagaat gcgacccact gcttccagtg agatcatggt cctacattgt agaaacacca      300

aactctgaga atggaatatg ttatccagga gatttcatcg actatgagga gctgagggag      360

caattgagct cagtgtcatc attcgaaaga ttcgaaatat ttcccaaaga aagctcatgg      420

cccaaccaca acacaaacgg agtaacggca gcatgctccc atgaggggaa aagcagtttt      480

tacagaaatt tgctatggct gacggagaag gagggctcat acccaaagct gaaaaattct      540

tatgtgaaca aaaaagggaa agaagtcctt gtactgtggg gtattcatca cccgcctaac      600

agtaaggaac aacagaatct ctatcagaat gaaaatgctt atgtctctgt agtgacttca      660

aattataaca ggagatttac cccggaaata gcagaaagac ccaaagtaag agatcaagct      720

gggaggatga actattactg gaccttgcta aaacccggag acacaataat atttgaggca      780

aatggaaatc taatagcacc aatgtatgct ttcgcactga gtagaggctt tgggtccggc      840

atcatcacct caaacgcatc aatgcatgag tgtaacacga agtgtcaaac acccctggga      900

gctataaaca gcagtctccc ttaccagaat atacacccag tcacaatagg agagtgccca      960

aaatacgtca ggagtgccaa attgaggatg gttacaggac taaggaacac tccgtccatt     1020

caatccagag gtctatttgg agccattgcc ggttttattg aagggggatg gactggaatg     1080

atagatggat ggtatggtta tcatcatcag aatgaacagg gatcaggcta tgcagcggat     1140

caaaaaagca cacaaaatgc cattaacggg attacaaaca aggtgaacac tgttatcgag     1200

aaaatgaaca ttcaattcac agctgtgggt aaagaattca acaaattaga aaaaaggatg     1260

gaaaatttaa ataaaaaagt tgatgatgga tttctggaca tttggacata taatgcagaa     1320

ttgttagttc tactggaaaa tgaaaggact ctggatttcc atgactcaaa tgtgaagaat     1380

ctgtatgaga aagtaaaaag ccaattaaag aataatgcca aagaaatcgg aaatggatgt     1440

tttgagttct accacaagtg tgacaatgaa tgcatggaaa gtgtaagaaa tgggacttat     1500

gattatccca aatattcaga agagtcaaag ttgaacaggg aaaaggtaga tggagtgaaa     1560

ttggaatcaa tggggatcta tcagattctg gcgatctact caactgtcgc cagttcactg     1620

gtgcttttgg tctccctggg ggcaatcagt ttctggatgt gttctaatgg atctttgcag     1680

tgcagaatat gcatctga                                                   1698


<210>  9
<211>  45
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - viral 3' UTR

<400>  9
gattagaatt tcagaaatat gaggaaaaac acccttgttt ctact                       45


<210>  10
<211>  40
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - portion of a neuraminidase (NA) protein comprising 
       the transmembrane domain and cytoplasmic tail

<400>  10

Met Asn Pro Asn Gln Lys Ile Thr Thr Ile Gly Ser Ile Cys Leu Val 
1               5                   10                  15      


Val Gly Leu Ile Ser Leu Ile Leu Gln Ile Gly Asn Ile Ile Ser Ile 
            20                  25                  30          


Trp Ile Ser His Ser Ile Gln Thr 
        35                  40  


<210>  11
<211>  223
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - receptor binding domain (RBD) of the spike protein 
       from the SARS-CoV-2 Wuhan variant

<400>  11

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn 
1               5                   10                  15      


Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val 
            20                  25                  30          


Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser 
        35                  40                  45              


Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val 
    50                  55                  60                  


Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp 
65                  70                  75                  80  


Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln 
                85                  90                  95      


Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr 
            100                 105                 110         


Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly 
        115                 120                 125             


Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys 
    130                 135                 140                 


Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr 
145                 150                 155                 160 


Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser 
                165                 170                 175     


Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val 
            180                 185                 190         


Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly 
        195                 200                 205             


Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 
    210                 215                 220             


<210>  12
<211>  11
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - flag tag

<400>  12

Gly Ser Gly Asp Tyr Lys Asp Asp Asp Asp Lys 
1               5                   10      


<210>  13
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - 2A self-cleaving peptide

<400>  13

Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val 
1               5                   10                  15      


Glu Glu Asn Pro Gly Pro 
            20          


<210>  14
<211>  565
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - influenza hemagglutinin (HA) protein

<400>  14

Met Lys Ala Asn Leu Leu Val Leu Leu Ser Ala Leu Ala Ala Ala Asp 
1               5                   10                  15      


Ala Asp Thr Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr 
            20                  25                  30          


Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn 
        35                  40                  45              


Leu Leu Glu Asp Ser His Asn Gly Lys Leu Cys Arg Leu Lys Gly Ile 
    50                  55                  60                  


Ala Pro Leu Gln Leu Gly Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly 
65                  70                  75                  80  


Asn Pro Glu Cys Asp Pro Leu Leu Pro Val Arg Ser Trp Ser Tyr Ile 
                85                  90                  95      


Val Glu Thr Pro Asn Ser Glu Asn Gly Ile Cys Tyr Pro Gly Asp Phe 
            100                 105                 110         


Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe 
        115                 120                 125             


Glu Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn His Asn 
    130                 135                 140                 


Thr Asn Gly Val Thr Ala Ala Cys Ser His Glu Gly Lys Ser Ser Phe 
145                 150                 155                 160 


Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Glu Gly Ser Tyr Pro Lys 
                165                 170                 175     


Leu Lys Asn Ser Tyr Val Asn Lys Lys Gly Lys Glu Val Leu Val Leu 
            180                 185                 190         


Trp Gly Ile His His Pro Pro Asn Ser Lys Glu Gln Gln Asn Leu Tyr 
        195                 200                 205             


Gln Asn Glu Asn Ala Tyr Val Ser Val Val Thr Ser Asn Tyr Asn Arg 
    210                 215                 220                 


Arg Phe Thr Pro Glu Ile Ala Glu Arg Pro Lys Val Arg Asp Gln Ala 
225                 230                 235                 240 


Gly Arg Met Asn Tyr Tyr Trp Thr Leu Leu Lys Pro Gly Asp Thr Ile 
                245                 250                 255     


Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Met Tyr Ala Phe Ala 
            260                 265                 270         


Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala Ser Met 
        275                 280                 285             


His Glu Cys Asn Thr Lys Cys Gln Thr Pro Leu Gly Ala Ile Asn Ser 
    290                 295                 300                 


Ser Leu Pro Tyr Gln Asn Ile His Pro Val Thr Ile Gly Glu Cys Pro 
305                 310                 315                 320 


Lys Tyr Val Arg Ser Ala Lys Leu Arg Met Val Thr Gly Leu Arg Asn 
                325                 330                 335     


Thr Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe 
            340                 345                 350         


Ile Glu Gly Gly Trp Thr Gly Met Ile Asp Gly Trp Tyr Gly Tyr His 
        355                 360                 365             


His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Gln Lys Ser Thr 
    370                 375                 380                 


Gln Asn Ala Ile Asn Gly Ile Thr Asn Lys Val Asn Thr Val Ile Glu 
385                 390                 395                 400 


Lys Met Asn Ile Gln Phe Thr Ala Val Gly Lys Glu Phe Asn Lys Leu 
                405                 410                 415     


Glu Lys Arg Met Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe Leu 
            420                 425                 430         


Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn Glu 
        435                 440                 445             


Arg Thr Leu Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Glu Lys 
    450                 455                 460                 


Val Lys Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly Cys 
465                 470                 475                 480 


Phe Glu Phe Tyr His Lys Cys Asp Asn Glu Cys Met Glu Ser Val Arg 
                485                 490                 495     


Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ser Lys Leu Asn 
            500                 505                 510         


Arg Glu Lys Val Asp Gly Val Lys Leu Glu Ser Met Gly Ile Tyr Gln 
        515                 520                 525             


Ile Leu Ala Ile Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Leu Val 
    530                 535                 540                 


Ser Leu Gly Ala Ile Ser Phe Trp Met Cys Ser Asn Gly Ser Leu Gln 
545                 550                 555                 560 


Cys Arg Ile Cys Ile 
                565 


<210>  15
<211>  1273
<212>  PRT
<213>  SARS-CoV-2

<400>  15

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val 
1               5                   10                  15      


Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe 
            20                  25                  30          


Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu 
        35                  40                  45              


His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp 
    50                  55                  60                  


Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp 
65                  70                  75                  80  


Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu 
                85                  90                  95      


Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser 
            100                 105                 110         


Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile 
        115                 120                 125             


Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr 
    130                 135                 140                 


Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr 
145                 150                 155                 160 


Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu 
                165                 170                 175     


Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe 
            180                 185                 190         


Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr 
        195                 200                 205             


Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu 
    210                 215                 220                 


Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr 
225                 230                 235                 240 


Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser 
                245                 250                 255     


Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro 
            260                 265                 270         


Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala 
        275                 280                 285             


Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys 
    290                 295                 300                 


Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val 
305                 310                 315                 320 


Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys 
                325                 330                 335     


Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala 
            340                 345                 350         


Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu 
        355                 360                 365             


Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro 
    370                 375                 380                 


Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe 
385                 390                 395                 400 


Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly 
                405                 410                 415     


Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys 
            420                 425                 430         


Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn 
        435                 440                 445             


Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe 
    450                 455                 460                 


Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys 
465                 470                 475                 480 


Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly 
                485                 490                 495     


Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val 
            500                 505                 510         


Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys 
        515                 520                 525             


Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn 
    530                 535                 540                 


Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu 
545                 550                 555                 560 


Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val 
                565                 570                 575     


Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe 
            580                 585                 590         


Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val 
        595                 600                 605             


Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile 
    610                 615                 620                 


His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser 
625                 630                 635                 640 


Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val 
                645                 650                 655     


Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala 
            660                 665                 670         


Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala 
        675                 680                 685             


Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 
    690                 695                 700                 


Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 
705                 710                 715                 720 


Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 
                725                 730                 735     


Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 
            740                 745                 750         


Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 
        755                 760                 765             


Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 
    770                 775                 780                 


Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 
785                 790                 795                 800 


Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 
                805                 810                 815     


Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 
            820                 825                 830         


Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 
        835                 840                 845             


Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 
    850                 855                 860                 


Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 
865                 870                 875                 880 


Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 
                885                 890                 895     


Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 
            900                 905                 910         


Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 
        915                 920                 925             


Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 
    930                 935                 940                 


Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 
945                 950                 955                 960 


Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 
                965                 970                 975     


Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 
            980                 985                 990         


Ile Asp Arg Leu Ile Thr Gly Arg  Leu Gln Ser Leu Gln  Thr Tyr Val 
        995                 1000                 1005             


Thr Gln  Gln Leu Ile Arg Ala  Ala Glu Ile Arg Ala  Ser Ala Asn 
    1010                 1015                 1020             


Leu Ala  Ala Thr Lys Met Ser  Glu Cys Val Leu Gly  Gln Ser Lys 
    1025                 1030                 1035             


Arg Val  Asp Phe Cys Gly Lys  Gly Tyr His Leu Met  Ser Phe Pro 
    1040                 1045                 1050             


Gln Ser  Ala Pro His Gly Val  Val Phe Leu His Val  Thr Tyr Val 
    1055                 1060                 1065             


Pro Ala  Gln Glu Lys Asn Phe  Thr Thr Ala Pro Ala  Ile Cys His 
    1070                 1075                 1080             


Asp Gly  Lys Ala His Phe Pro  Arg Glu Gly Val Phe  Val Ser Asn 
    1085                 1090                 1095             


Gly Thr  His Trp Phe Val Thr  Gln Arg Asn Phe Tyr  Glu Pro Gln 
    1100                 1105                 1110             


Ile Ile  Thr Thr Asp Asn Thr  Phe Val Ser Gly Asn  Cys Asp Val 
    1115                 1120                 1125             


Val Ile  Gly Ile Val Asn Asn  Thr Val Tyr Asp Pro  Leu Gln Pro 
    1130                 1135                 1140             


Glu Leu  Asp Ser Phe Lys Glu  Glu Leu Asp Lys Tyr  Phe Lys Asn 
    1145                 1150                 1155             


His Thr  Ser Pro Asp Val Asp  Leu Gly Asp Ile Ser  Gly Ile Asn 
    1160                 1165                 1170             


Ala Ser  Val Val Asn Ile Gln  Lys Glu Ile Asp Arg  Leu Asn Glu 
    1175                 1180                 1185             


Val Ala  Lys Asn Leu Asn Glu  Ser Leu Ile Asp Leu  Gln Glu Leu 
    1190                 1195                 1200             


Gly Lys  Tyr Glu Gln Tyr Ile  Lys Trp Pro Trp Tyr  Ile Trp Leu 
    1205                 1210                 1215             


Gly Phe  Ile Ala Gly Leu Ile  Ala Ile Val Met Val  Thr Ile Met 
    1220                 1225                 1230             


Leu Cys  Cys Met Thr Ser Cys  Cys Ser Cys Leu Lys  Gly Cys Cys 
    1235                 1240                 1245             


Ser Cys  Gly Ser Cys Cys Lys  Phe Asp Glu Asp Asp  Ser Glu Pro 
    1250                 1255                 1260             


Val Leu  Lys Gly Val Lys Leu  His Tyr Thr 
    1265                 1270             


<210>  16
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - RT-PCR primer

<400>  16
gtagatgcag caaaagcagg ggaaaata                                          28


<210>  17
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - RT-PCR primer

<400>  17
ccatcctcaa tttggcac                                                     18


<210>  18
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - RT-PCR primer

<400>  18
agcaaaagca ggggaaaata                                                   20


<210>  19
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic - RT-PCR primer

<400>  19
gtcttcgagc aggttaacag                                                   20


<210>  20
<211>  223
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - receptor binding domain (RBD) of the spike protein 
       from the SARS-CoV-2 Beta variant

<400>  20

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn 
1               5                   10                  15      


Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val 
            20                  25                  30          


Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser 
        35                  40                  45              


Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val 
    50                  55                  60                  


Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp 
65                  70                  75                  80  


Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln 
                85                  90                  95      


Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr 
            100                 105                 110         


Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly 
        115                 120                 125             


Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys 
    130                 135                 140                 


Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr 
145                 150                 155                 160 


Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser 
                165                 170                 175     


Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val 
            180                 185                 190         


Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly 
        195                 200                 205             


Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 
    210                 215                 220             


<210>  21
<211>  223
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - receptor binding domain (RBD) of the spike protein 
       from the SARS-CoV-2 Gamma variant

<400>  21

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn 
1               5                   10                  15      


Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val 
            20                  25                  30          


Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser 
        35                  40                  45              


Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val 
    50                  55                  60                  


Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp 
65                  70                  75                  80  


Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln 
                85                  90                  95      


Thr Gly Thr Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr 
            100                 105                 110         


Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly 
        115                 120                 125             


Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys 
    130                 135                 140                 


Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr 
145                 150                 155                 160 


Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser 
                165                 170                 175     


Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val 
            180                 185                 190         


Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly 
        195                 200                 205             


Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 
    210                 215                 220             


<210>  22
<211>  223
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - receptor binding domain (RBD) of the spike protein 
       from the SARS-CoV-2 Delta variant

<400>  22

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn 
1               5                   10                  15      


Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val 
            20                  25                  30          


Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser 
        35                  40                  45              


Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val 
    50                  55                  60                  


Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp 
65                  70                  75                  80  


Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln 
                85                  90                  95      


Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr 
            100                 105                 110         


Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly 
        115                 120                 125             


Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys 
    130                 135                 140                 


Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys 
145                 150                 155                 160 


Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser 
                165                 170                 175     


Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val 
            180                 185                 190         


Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly 
        195                 200                 205             


Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 
    210                 215                 220             


<210>  23
<211>  223
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic - receptor binding domain (RBD) of the spike protein 
       from the SARS-CoV-2 Omicron variant

<400>  23

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn 
1               5                   10                  15      


Leu Cys Pro Phe Asp Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val 
            20                  25                  30          


Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser 
        35                  40                  45              


Val Leu Tyr Asn Leu Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val 
    50                  55                  60                  


Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp 
65                  70                  75                  80  


Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln 
                85                  90                  95      


Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr 
            100                 105                 110         


Gly Cys Val Ile Ala Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser 
        115                 120                 125             


Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys 
    130                 135                 140                 


Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys 
145                 150                 155                 160 


Pro Cys Asn Gly Val Ala Gly Phe Asn Cys Tyr Phe Pro Leu Lys Ser 
                165                 170                 175     


Tyr Ser Phe Arg Pro Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val 
            180                 185                 190         


Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly 
        195                 200                 205             


Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 
    210                 215                 220             


